title: DNA polymerase κ-dependent DNA synthesis at stalled replication forks is important for CHK1 activation.
abstract: Formation of primed single-stranded DNA at stalled replication forks triggers activation of the replication checkpoint signalling cascade resulting in the ATR-mediated phosphorylation of the Chk1 protein kinase, thus preventing genomic instability. By using siRNA-mediated depletion in human cells and immunodepletion and reconstitution experiments in Xenopus egg extracts, we report that the Y-family translesion (TLS) DNA polymerase kappa (Pol κ) contributes to the replication checkpoint response and is required for recovery after replication stress. We found that Pol κ is implicated in the synthesis of short DNA intermediates at stalled forks, facilitating the recruitment of the 9-1-1 checkpoint clamp. Furthermore, we show that Pol κ interacts with the Rad9 subunit of the 9-1-1 complex. Finally, we show that this novel checkpoint function of Pol κ is required for the maintenance of genomic stability and cell proliferation in unstressed human cells.
authors.affiliation: Equipe Labellisée La Ligue Contre le Cancer 2013, INSERM UMR 1037, CNRS ERL 505294, CRCT (Cancer Research Center of Toulouse), Toulouse, France.
authors.forename: Rémy
authors.initials: R
authors.lastname: Bétous
authors.forename: Marie-Jeanne
authors.initials: MJ
authors.lastname: Pillaire
authors.forename: Laura
authors.initials: L
authors.lastname: Pierini
authors.forename: Siem
authors.initials: S
authors.lastname: van der Laan
authors.forename: Bénédicte
authors.initials: B
authors.lastname: Recolin
authors.forename: Emma
authors.initials: E
authors.lastname: Ohl-Séguy
authors.forename: Caixia
authors.initials: C
authors.lastname: Guo
authors.forename: Naoko
authors.initials: N
authors.lastname: Niimi
authors.forename: Petr
authors.initials: P
authors.lastname: Grúz
authors.forename: Takehiko
authors.initials: T
authors.lastname: Nohmi
authors.forename: Errol
authors.initials: E
authors.lastname: Friedberg
authors.forename: Christophe
authors.initials: C
authors.lastname: Cazaux
authors.forename: Domenico
authors.initials: D
authors.lastname: Maiorano
authors.forename: Jean-Sébastien
authors.initials: JS
authors.lastname: Hoffmann
mesh_terms: D000818:Animals; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D049109:Cell Proliferation; D000071877:Checkpoint Kinase 1; D004261:DNA Replication / Q000502:physiology*; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D042822:Genomic Instability / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D029867:Xenopus Proteins / Q000235:genetics / Q000378:metabolism*; D014982:Xenopus laevis; D000099270:Y-Family DNA Polymerases
chemical_list: D018797:Cell Cycle Proteins; D000071877:Checkpoint Kinase 1; D004259:DNA-Directed DNA Polymerase; D011494:Protein Kinases; D029867:Xenopus Proteins; C069930:rad9 protein; C000605867:CHEK1 protein, human; C000605875:Chek1 protein, Xenopus; C404302:POLK protein, human; D000099270:Y-Family DNA Polymerases; C530191:Polk protein, Xenopus